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Development and evaluation of strain typing methods for Mycobacterium bovis - SE3017

Description
Despite the current test and slaughter policy to control bovine tuberculosis in cattle, the prevalence of bovine tuberculosis is increasing, particularly in the south-west of England. The increasing incidence is thought to be associated with transmission from infected badgers. However, although the sum of evidence strongly supports this view, the Krebs Report recommends that there is a need for a better understanding of herd breakdowns. In light of this recommendation, MAFF has highlighted a requirement ‘to develop sensitive and specific molecular typing techniques to enable a long-term study of transmission between wildlife, cattle and other species (including man) and to assess the variation of different genomes of M. bovis’. The aim of this proposal is to evaluate the suitability of all current molecular typing techniques (REA, RFLP, spoligotyping and VNTR) for the GB situation and to develop more sensitive methods which allow high-throughput molecular typing (AFLP, Affymetrix chip technology, improved RFLP and VNTR and blood-based fingerprinting techniques). These new methods will be validated against existing methods using epidemiologically defined M. bovis isolates from GB. We also aim to standardise fingerprinting techniques between VLA and PHLS to allow co-ordinated surveillance of Mycobacterium bovis infection in animals and humans in Great Britain.
Objective
Obj. No. Completed by date Description
01
VLA 01/01/2002 To culture and distribute 200 GB M. bovis isolates to collaborating laboratories
02
QUB 01/08/2002 To apply RFLP (PGRS) to 200 M. bovis GB isolates
03
AgR 01/08/2002 To apply REA to 200 M. bovis GB isolates
04
VLA 01/01/2003 To apply fluorescent VNTR to 200 M. bovis GB isolates
05
VLA
PHLS 01/02/2002 To standardise and validate methodology of RFLP, spoligotyping and VNTR between VLA and PHLS using 50 human isolates
06
QUB 01/05/2002 To identify new targets for RFLP (QUB) and apply to 50 selected isolates
07
VLA
QUB 01/11/2002 To identify new targets for VNTR and apply to 50 selected isolates
07a
VLA 01/02/2003 To develop fluorescent AFLP for M. bovis and apply to the 200 M. bovis GB isolates
08
RIVM 01/08/2003 To assess feasibility of using microarray technology for typing M. bovis using the 200 M. bovis GB isolates
09
PHLS 01/09/2002 To determine DNA sequence of DR region of M. bovis from 50 GB human TB M. bovis isolates
10
IC 31/08/2000 To develop a genotypic typing system based on variations in PE/PPE genes
11
IC 30/03/2002 To determine whether expression of PE/PPE genes result in a different antigenic phenotype
12
IC 01/08/2002 To apply genotypic and phenotypic assays to monitor strain variation of M. bovis in badger and cattle populations
13
PHLS 31/12/2001 To collect M. bovis-positive human sera for evaluation of antigenic fingerprinting
14 31/08/2003 To evaluate results and prepare document of results and recommendations
15 31/03/2003 To apply VNTR to isolates collected during 2000 and to badger trial samples
Project Documents
• Final Report : Development and evaluation of strain typing methods for M. bovis   (252k)
Time-Scale and Cost
From: 1999

To: 2005

Cost: £1,275,223
Contractor / Funded Organisations
Veterinary Laboratories Agency
Keywords
Animal Health              
Biotechnology              
Bovine Tuberculosis              
Control              
Diagnosis              
Plants and Animals              
Zoonoses              
Fields of Study
Animal Health